RESUMO
It is increasingly common in oncology practice to perform tumour sequencing using large cancer panels. For pathogenic sequence variants in cancer susceptibility genes identified on tumour-only sequencing, it is often unclear whether they are of somatic or constitutional (germline) origin. There is wide-spread disparity regarding both the extent to which systematic 'germline-focussed analysis' is carried out upon tumour sequencing data and for which variants follow-up analysis of a germline sample is carried out. Here we present analyses of paired sequencing data from 17 152 cancer samples, in which 1494 pathogenic sequence variants were identified across 65 cancer susceptibility genes. From these analyses, the European Society of Medical Oncology Precision Medicine Working Group Germline Subgroup has generated (i) recommendations regarding germline-focussed analyses of tumour-only sequencing data, (ii) indications for germline follow-up testing and (iii) guidance on patient information-giving and consent.
Assuntos
Biomarcadores Tumorais/genética , Testes Genéticos/normas , Neoplasias/diagnóstico , Medicina de Precisão/métodos , Análise Mutacional de DNA , União Europeia , Predisposição Genética para Doença , Mutação em Linhagem Germinativa , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Consentimento Livre e Esclarecido/normas , Oncologia/métodos , Oncologia/normas , Neoplasias/genética , Guias de Prática Clínica como Assunto , Medicina de Precisão/normas , Sociedades Médicas/normasAssuntos
Doença Celíaca/diagnóstico , Doença Celíaca/epidemiologia , Fraturas Ósseas/diagnóstico , Fraturas Ósseas/epidemiologia , Programas de Rastreamento/métodos , Admissão do Paciente , Idoso , Idoso de 80 Anos ou mais , Doença Celíaca/sangue , Estudos de Coortes , Feminino , Fraturas Ósseas/sangue , Humanos , Pacientes Internados , Masculino , Programas de Rastreamento/normas , Osteoporose/sangue , Osteoporose/diagnóstico , Osteoporose/epidemiologia , Admissão do Paciente/normas , Estudos RetrospectivosRESUMO
In many parts of the nervous system, neurons with the same function often have similar "birth dates" (the time their precursor withdrew from the cell cycle). We investigated the birth dates of eight functional classes of rat sympathetic postganglionic neurons by injecting bromodeoxyuridine during embryonic development, while retrograde tracing and immunohistochemistry were used to identify postganglionic neurons of different functional classes in the mature animals. The times of withdrawal from the cell cycle overlapped, but there were significant differences in the peak time of withdrawal for most of the classes. Furthermore, sympathetic cholinergic postganglionic neurons had a significantly greater proportion of their total population labelled with bromodeoxyuridine than did any of the noradrenergic classes of neurons, indicating prenatal class-specific differences in the handling of bromodeoxyuridine. Together, our findings indicate that, prior to extending axons to their targets, different functional classes of sympathetic neurons show differences in phenotype.
Assuntos
Sistema Nervoso Autônomo/metabolismo , Gânglios Simpáticos/metabolismo , Neurônios/metabolismo , Animais , Bromodesoxiuridina/farmacologia , Diferenciação Celular , Biologia do Desenvolvimento/métodos , Regulação da Expressão Gênica no Desenvolvimento , Imuno-Histoquímica/métodos , Neuropeptídeos/química , Fenótipo , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
A number of 3-alkyl analogues of the experimental antitumor drug mitozolomide [8-carbamoyl-3-(2-chloroethyl)imidazo[5,1-d]-1,2,3,5-tetrazin-4(3H )-one] have been screened against murine tumors in vivo. Only the compounds with a 3-methyl- or 3-bromoethyl group possessed significant antitumor activity against the TLX5 lymphoma. The 3-methyl analogue, 8-carbamoyl-3-methylimidazo[5,1-d]-1,2,3,5-tetrazin-4(3H)-one (CCRG 81045), was investigated further and found to possess good activity, when administered i.p., against the L1210 and P388 leukemias, the M5076 reticulum cell sarcoma, B16 melanoma, and ADJ/PC6A plasmacytoma. The drug was also active when administered p.o. to mice bearing the L1210 leukemia. A daily for 5 days schedule of 100 mg/kg CCRG 81045 produced increases of survival time of treated animals compared to controls of 176 and greater than 235% against the P388 and L1210 leukemias, respectively. In the female C57BL x DBA/2 F1 mouse the 10% lethal dose was 125 mg/kg daily for 5 days. CCRG 81045 was found to undergo mild alkaline hydrolysis and ring fission to form the linear triazene 5-(3-methyltriazen-1-yl)imidazole-4-carboxamide, which is the putative metabolite formed upon metabolic activation of the antitumor drug dacarbazine [5-(3,3-dimethyltriazen-1-yl)imidazole-4-carboxamide]. The half-life of CCRG 81045 at 37 degrees C in 0.2 M phosphate buffer (pH 7.4) was 1.24 h, whereas that of 5-(3-methyltriazen-1-yl)imidazole-4-carboxamide at 25 degrees C was reported to be 8 min (F. H. Shealy and C. A. Krauth, J. Med. Chem., 9:34-37, 1966). The half-life of CCRG 81045 in human plasma in vitro at 37 degrees C was 0.42 h. Pharmacokinetic experiments conducted in BALB/c mice produced plasma profiles of CCRG 81045, administered i.p. or p.o., which showed a rapid absorption phase, elimination half-lives of 1.13 h (i.p.) and 1.29 h (p.o.), and a bioavailability of 0.98.
Assuntos
Antineoplásicos/uso terapêutico , Dacarbazina/uso terapêutico , Imidazóis/uso terapêutico , Neoplasias Experimentais/tratamento farmacológico , Animais , Imidazóis/farmacocinética , Imidazóis/toxicidade , Neoplasias Pulmonares/tratamento farmacológico , Masculino , Melanoma Experimental/tratamento farmacológico , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos CBA , Camundongos Endogâmicos DBA , Camundongos Endogâmicos , Plasmocitoma/tratamento farmacológico , Sarcoma Experimental/tratamento farmacológico , Especificidade da Espécie , TemozolomidaRESUMO
A series of derivatives of N-methylformamide (NMF), an experimental antitumor agent, has been prepared, having the general formula R3C(X)NR1R2 where R1 = H, CH3, CD3, CH2CF3, CH2CH2Cl, cyclopropyl, C2H5, CH2OH, CH2OR, CH2N(CH3)2; R2 = H, CH3; R3 = H, CF3, CCl3, CH3, Ph, NHCH3, N(CH3)2; and X = O, S, NH. A further short series of "push-pull" olefins of the general formula R1R2C = CHNR3R4 has been synthesized where R1 = H, CH3 and R2 = H, NO2, CN, CHO, CH3 and R3 = H and R4 = H, CH3, morpholino. These compounds have been tested for activity against the M5076 ovarian sarcoma and the TLX5 lymphoma in mice. NMF was by far the most potent agent of both series with activity against both tumors. Some other compounds showed weak activity, but there is a rigorous structural requirement for activity and most analogues were inactive. Certain members of the series exist as equilibrium mixtures of rotamers about the amide or pro-amide bonds as shown by NMR.
Assuntos
Antineoplásicos/farmacologia , Formamidas/farmacologia , Animais , Feminino , Formamidas/metabolismo , Hidroxilação , Linfoma/tratamento farmacológico , Espectroscopia de Ressonância Magnética , Camundongos , Neoplasias Ovarianas/tratamento farmacológico , Relação Estrutura-AtividadeRESUMO
Aspects of the toxicology of N-methylformamide (NMF), an investigational antitumour agent, were studied in mice. After injection of NMF at its LD10 (800 mg/kg) dosage the total peripheral white blood cell and platelet counts were unchanged in BALB/c mice. A mild granulocytosis was seen in this strain after administration of the LD50 (2300 mg/kg) dosage. Plasma activity of the enzyme sorbitol dehydrogenase in BDF1 mice was markedly increased after either a single injection of not less than 800 mg/kg or a chronic treatment of not less than 400 mg/kg/day over 5 days indicating the drug to be hepatotoxic. Plasma activities of L-alanine and L-aspartate aminotransferases were also increased after the chronic treatment. Chronic administration of NMF was less hepatotoxic than single dose administration of the same total dose and also increased the antitumour efficacy of NMF against the M5076 sarcoma. These results indicate that the maximum therapeutic benefit of NMF might be obtained by the use of chronic schedules and that the drug is not myelosuppressive.
Assuntos
Formamidas/toxicidade , Alanina Transaminase/sangue , Animais , Antineoplásicos , Aspartato Aminotransferases/sangue , Ciclofosfamida/toxicidade , Formamidas/uso terapêutico , Granulócitos/citologia , L-Iditol 2-Desidrogenase/sangue , Dose Letal Mediana , Contagem de Leucócitos , Fígado/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Contagem de Plaquetas , Sarcoma Experimental/tratamento farmacológicoRESUMO
The use of contrast lymphangiography is a relatively new technique in lymphatic anatomical research, employed as a tool for evaluating lymphatic anatomy in fresh cadaveric specimens. With the use of microsurgical techniques to cannulate lymphatics, contrast media can enable the use of lymphangiography for evaluating lymphatic anatomy. However, the ability to cannulate lymphatics with diameters that are less than the smallest commercially available needles has been a significant limitation, and indeed a challenge. The smallest commercially available hypodermic needles have been 30-gauge needles, with 0.3 mm outer diameters. The lymphatics for cannulation in our studies are of the order of 0.1 mm, and other options have been required. We describe a novel technique for cannulating lymphatic vessels, creating a modified glass hypodermic needle. We have shown that these glass needles can be made with accuracy to diameters as low as 0.01 mm. Although 0.1 mm glass needles are the more commonly utilized in most dissections, we can now accurately create these glass needles to any caliber between 0.01 mm and 0.1 mm, based on the predicted lymphatic anatomy.
Assuntos
Meios de Contraste/administração & dosagem , Vasos Linfáticos/anatomia & histologia , Vasos Linfáticos/diagnóstico por imagem , Linfografia/instrumentação , Agulhas , Cadáver , Meios de Contraste/farmacologia , Desenho de Equipamento , Vidro , Temperatura Alta , Humanos , Sistema Linfático/anatomia & histologia , Linfografia/métodosAssuntos
Transplante Ósseo/patologia , Fáscia/transplante , Monitorização Fisiológica/instrumentação , Músculo Esquelético/transplante , Transplante de Pele/patologia , Retalhos Cirúrgicos/irrigação sanguínea , Fáscia/irrigação sanguínea , Humanos , Fluxometria por Laser-Doppler , Músculo Esquelético/irrigação sanguínea , Complicações Pós-Operatórias/prevenção & controle , Reoperação , Espectrofotometria , Resultado do TratamentoRESUMO
1. The effect of an infusion of sodium beta-hydroxybutyrate on leucine and alanine metabolism was investigated in dogs starved for 12 h. To determine whether the metabolic changes produced by this infusion were due to the resultant alkalaemia the effect of an equimolar infusion of sodium bicarbonate was also studied. 2. The sodium beta-hydroxybutyrate infusion reduced alanine concentration as a result of a decrease in alanine production rate and an increase in alanine metabolic clearance rate. The sodium bicarbonate infusion induced a small decrease in alanine concentration which was due to an increased metabolic clearance rate. Alanine production rate showed no change. This demonstrates that the fall in alanine concentration after a sodium beta-hydroxybutyrate infusion is due both to a ketone-specific inhibitory effect on alanine production rate and an increased metabolic clearance rate caused by the alkalaemia. 3. Leucine concentration was increased after the ketone infusion due to a small increase in production rate and there was a small increase in the rate of leucine incorporation into protein. Alkalaemia had no effect on leucine concentration or metabolism.
Assuntos
Alanina/metabolismo , Hidroxibutiratos/farmacologia , Corpos Cetônicos/metabolismo , Leucina/metabolismo , Ácido 3-Hidroxibutírico , Animais , Bicarbonatos/farmacologia , Cães , Taxa de Depuração Metabólica , Biossíntese de Proteínas , Sódio/farmacologia , Bicarbonato de Sódio , Inanição/metabolismoRESUMO
N-Methylformamide (NMF) was found to be non-toxic to the bone marrow as reflected in the absence of leukopenia in mice, even when the marrow had been compromised by prior administration of cyclophosphamide. Thus recovery from the leukopenic nadir after 160 mg/kg of cyclophosphamide was unaffected by 200 mg/kg X 10 of NMF. This combination, given to animals bearing the M5076 sarcoma, proved to have an additive antitumour effect as measured by tumour growth delay and was superior to the antitumour effect of two doses of cyclophosphamide, a regime which prolonged the leukopenia. Furthermore, the hepatotoxicity of NMF was not augmented by the addition of cyclophosphamide. When hepatotoxicity was induced in BALB/c mice bearing the NMF-resistant ADJ/PC6A plasmacytoma, cyclophosphamide fully maintained its antitumour effect. The results show NMF to be a highly specific antiproliferative agent with potential for use in the therapy of patients with a compromised bone marrow and/or in combination chemotherapy.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Formamidas/administração & dosagem , Sarcoma Experimental/tratamento farmacológico , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Peso Corporal/efeitos dos fármacos , Medula Óssea/efeitos dos fármacos , Doença Hepática Induzida por Substâncias e Drogas , Ciclofosfamida/administração & dosagem , Ciclofosfamida/efeitos adversos , Feminino , Formamidas/efeitos adversos , Contagem de Leucócitos , Camundongos , Camundongos Endogâmicos , Plasmocitoma/tratamento farmacológico , Sarcoma Experimental/patologiaRESUMO
A series of derivatives of the anti-tumour hydroxymethyltriazenes have been investigated for activity in vivo and in vitro. Acetoxymethyltriazenes are active in vivo against the TLX5, P388 and PC6 tumours in mice, and inhibit the growth of TLX5, Np and Li cells in vitro without metabolic activation. The acetoxymethyltriazenes are comparable with the hydroxymethyltriazenes and monomethyltriazenes in their spectrum of activity and thus appear to be prodrugs for these species. On the other hand, a methoxymethyltriazene was found to be active on the TLX5 tumour in vivo, but did not inhibit the growth of Np cells in vitro. This latter observation is consistent with the anticipated chemical stability of the methoxymethyltriazene and the requirement for metabolic O-demethylation to generate an active species. Acetoxymethyltriazenes do not require metabolic intervention and break down chemically in phosphate buffer to the hydroxymethyltriazene, which in turn loses formaldehyde to give the incipient methylating agent, the monomethyltriazene.
Assuntos
Antineoplásicos/farmacologia , Dacarbazina/metabolismo , Farmacologia , Pró-Fármacos/farmacologia , Triazenos/farmacologia , Animais , Estabilidade de Medicamentos , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos CBA , Neoplasias Experimentais/tratamento farmacológico , Triazenos/síntese química , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
Leucine and alanine production rate was measured in 5 patients with acid maltase deficiency in the postabsorptive state, following 6 months on a normal diet with placebo and 6 months on an isocaloric high protein diet (16-22% protein). Whole body leucine production rate, a measure of protein degradation, expressed in terms of lean body mass was significantly greater than in five control subjects. Following the high protein diet, leucine production rate was decreased in four of the five patients but this was not statistically significant. Alanine production rate expressed in terms of lean body mass was significantly greater than in control subjects. After the high protein diet, alanine production rate and concentration were significantly decreased (p less than 0.05). There were no significant improvements in any of the clinically relevant variables measured in these patients. It is possible that a larger increase in protein intake over a longer time period may have a clinical effect.